Page 152 - Advances in Forensic Applications of Mass Spectrometry - Jehuda Yinon

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sample preparation involved a decontamination step of the hair with meth-
ylene chloride and the sonication in methanol of 100 mg of powdered hair
for 2 h. After elimination of the solvent, the hair sample was solubilized in 1
ml 1 N NaOH, 15 min at 95˚C, in the presence of 10 ng stanozolol-d used
3
as internal standard. The homogenate was neutralized and extracted using
consecutively a solid phase (Isolute C18) and a liquid–liquid (pentane) extrac-
tion. After evaporation of the final organic phase, the dry extract was deriva-
tized using 40 ml MBHFA/TMSI (1000:20, v/v), incubated for 5 min at 80˚C,
followed by 10 ml of MBHFBA, incubated for 30 min at 80˚C. The derivatized
extract was analyzed by a Hewlett–Packard GC/MS system with a 5989 B
Engine operating in negative chemical ionization (NCI) mode of detection.
Figure 3.2 shows the mass spectrum of an HFB-derivative of stanozolol in
the NCI mode. The assay was able to detect 2 pg of stanozolol per mg of hair
when approximately 100 mg hair material were processed. The analysis of a
3-cm-long hair strand, obtained from a young bodybuilder (27 years old)
®
declaring to be a regular user of Winstrol (stanozolol, 2 mg), revealed the
presence of stanozolol at the concentration of 15 pg/mg (Figure 3.3).
More recently, a method for the quantitative determination of meth-
enolone in human hair was developed. The sample preparation involved a
12
decontamination step of the hair with methylene chloride. The hair sample
(about 100 mg) was solubilized in 1 ml NaOH 1 N, 15 min at 95˚C, in the
presence of 1 ng testosterone-d used as internal standard. The homogenate
3
was neutralized and extracted using consecutively a solid phase (Isolute C18
eluted with methanol) and a liquid–liquid (pentane) extraction. The residue
was derivatized by adding 50 ml MSTFA/NH I/2-mercaptoethanol (1000:2:5,
4
v/v/v), then incubated for 20 min at 60˚C. A 1.5-ml aliquot of the derivatized
extract was injected into the column (HP5-MS capillary column, 5% phenyl-
95% methylsiloxane, 30 m ¥ 0.25 mm i.d. ¥ 0.25 mm film thickness) of a
Hewlett–Packard (Palo Alto, CA) gas chromatograph (6890 Series). Meth-
enolone was detected by its parent ion at m/z 446 and daughter ions at m/z
208 and 195 by means of a Finnigan TSQ 700 MS/MS system. The assay was
capable of detecting 1 pg/mg of methenolone when approximately 100 mg

Figure 3.1 (See figure on facing page.) MS/MS chromatograms of an extract of
a human hair spiked with 50 pg/mg each of a series of drugs. Peaks: A = test-
osterone cypionate, sum of ions m/z 209 + 469, daughter ions of m/z 484; B =
testosterone phenyl propionate, sum of ions m/z 209 + 477, daughter ions of m/z
492; C = nandrolone decanoate, sum of ions m/z 182 + 194, daughter ions of m/z
500; D = testosterone decanoate, sum of ions m/z 209 + 499, daughter ions of
m/z 514; E = testosterone undecanoate, sum of ions m/z 209 + 513, daughter ions
of m/z 528. (Reprinted from Gaillard, Y. et al., J. Chromatogr. B, 735, 189, 1999.
Copyright 1999, with permission from Elsevier Science.)

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